基于纳米探针荧光成像技术在肾透明细胞癌治疗中的研究

Study on the treatment of clear cell renal cell carcinoma based on nanoprobe fluorescence imaging

目的为提高肾癌的术前诊断和治疗效果,研究制备一种可以靶向肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)的新型纳米探针(clear cell renal cell carcinoma targeting system,CTS),用以评估CTS在肾透明细胞癌荧光分子成像中的靶向性、荧光效应及其早期诊断价值。方法构建纳米探针CTS,分别与人肾透明细胞癌细胞系786-O和Caki-1共孵育,用激光共聚焦显微镜观察CTS与受体的靶向情况。然后建立黑鼠肾透明细胞癌皮下移植瘤模型,经小鼠尾静脉注射CTS后,进行活体荧光分子成像,观察CTS的成像效果,分析肿瘤部位的荧光信号强度。最后取小鼠肿瘤组织进行免疫组织化学技术(immunohistochemistry,IHC)和免疫荧光染色,检验CTS的靶向作用。结果激光共聚焦显微镜下观察发现,CTS与受体良好共定位。活体荧光分子成像结果显示,探针在实验鼠皮下肿瘤部位出现浓聚,显示CTS有较好的靶向标记性和信噪比。最后,取小鼠肿瘤组织进行IHC和冰冻切片免疫荧光染色,结果显示碳酸酐酶IX(carbonic anhydrase IX,CAIX)在ccRCC组织中过表达,并且CTS可以有效靶向CAIX。结论CTS具有很好的靶向ccRCC细胞的能力和成像效应,可通过荧光分子成像技术实现肿瘤早期诊断并指导肿瘤切除。

Objective To improve the preoperative diagnosis and treatment of renal cancer,a novel nanoprobes targeting clear cell renal cell carcinoma(ccRCC)clear cell renal cell carcinoma targeting system(CTS)is developed to evaluate the targeting ability,fluorescence effect,and early diagnostic value of CTS in fluorescence molecular imaging of clear cell renal cell carcinoma.Methods The nanoprobes CTS were constructed and co-cultured with human clear cell renal cell carcinoma cell lines 786-O and Caki-1.The targeting situation of CTS and the receptors was observed using a laser confocal microscope.Subsequently,a subcutaneous transplantation tumor model of clear cell renal cell carcinoma was established in black mice.After tail vein injection of CTS,in vivo fluorescence molecular imaging was performed to observe the imaging effect of CTS and analyze the fluorescence signal intensity at the tumor site.Finally,tumor tissues were collected from the mice for immunohistoghemistry(IHC)and immunofluorescence staining to examine the targeting effect of CTS.Results Under laser confocal microscopy,it was observed that CTS and the receptors co-localized well.The in vivo fluorescence molecular imaging results showed that the probe concentrated in the subcutaneous tumor site of the experimental mice,demonstrating that CTS had good targeting labeling and signal-to-noise ratio.Finally,tumor tissues were collected from the mice for IHC and frozen section immunofluorescence staining,and the results indicated that carbonic anhydrase IX(CAIX)was overexpressed in ccRCC tissues and that CTS could effectively target CAIX.Conclusion CTS has a good ability to target ccRCC cells and imaging effects,and it can achieve early tumor diagnosis and guide tumor resection through fluorescence molecular imaging technology.